Retention Time Table for Carbomix Phases

Retention time Table for Carbomix Phases

Compound Carbomix H-NP5 Carbomix H-NP10 Carbomix Ca-NP5 Carbomix Ca-NP10 Carbomix Pb-NP5 Carbomix Pb-NP10 8% 5% 8% 10% 8% 5% 8% 10% 8% 5% 8% Conditions: Acetic acid 13.58 17.52 14.64 13.39 / / / / / / / Carbomix H-NP10 Adonitol 9.85 13.59 11.10 10.26 13.40 16.76 14.73 13.67 18.81 21.88 19.88 Mobile phase：2.5 mM H2SO4 Arabinitol 10.06 13.82 11.30 10.41 15.99 19.80 17.72 16.06 23.63 27.76 24.95 Flow rate：0.6 mL/min D-(-)-Arabinose 9.77 13.46 10.93 10.08 12.13 13.77 13.43 12.52 14.81 17.74 15.76 Detector：RID L-(+)-Arabinose 9.77 13.45 10.93 10.08 12.13 15.45 13.41 12.53 14.82 17.74 15.76 ： 1,4-Butanediol 19.52 23.11 / 19.26 14.41 17.81 15.96 14.48 15.98 17.77 16.49 Column Tem. 80℃ n-Butanol 34.23 35.93 / 32.66 24.17 / / / 28.37 28.28 27.91 Injection Volume：10 μL (3 injections) t-Butanol 23.19 / / / 15.11 / / / 15.96 17.80 16.60 sec-butyl alcohol 28.55 31.90 30.25 28.23 18.82 22.83 21.06 19.01 20.63 22.25 21.06 Carbomix H-NP5 D-(+)-Cellobiose 7.79 9.79 8.18 8.17 8.42 10.10 8.81 8.96 10.30 12.27 10.89 Mobile phase：2.5 mM H2SO4 Citric acid 8.09 10.26 8.69 8.35 / / / / / / / Flow rate：0.6 mL/min Erythriyol 10.74 14.70 11.94 11.00 14.15 17.34 15.98 14.47 18.67 21.34 19.60 Detector：RID Ethanol 20.24 23.13 21.82 20.72 15.48 18.30 16.38 15.89 16.32 17.69 16.79 Column Tem.： 55℃ Formic acid 12.59 16.11 13.51 12.48 / / / / / / / Injection Volume：10 μL (3 injections) D-Fructose 9.25 12.65 10.27 9.58 12.08 15.27 13.34 12.45 15.85 19.17 16.86 Fumaric acid 11.71 / 13.90 / / / / / / / / Carbomix Ca-NP10 Galactitol 9.51 13.13 10.66 9.87 18.22 22.57 19.44 18.05 30.81 36.54 32.57 ： D-(+)-Galactose 9.13 12.54 10.15 9.48 10.73 13.77 11.77 11.20 13.54 16.50 14.52 Mobile phase H2O D-Glucose 8.76 11.90 9.68 9.16 9.82 12.37 10.61 10.32 11.97 14.31 12.74 Flow rate：0.6 mL/min Glycerol 12.19 16.07 13.61 12.46 14.96 17.89 16.20 15.12 18.34 20.59 19.13 Detector：RID IPA 21.94 25.79 23.89 22.53 15.37 18.61 17.16 15.88 16.27 17.86 16.78 Column Tem.： 85℃ Lactic acid 11.40 15.41 12.70 11.55 / / / / / / / Injection Volume：10 μL (3 injections) B-Lactose 7.93 10.24 8.42 8.29 8.70 10.81 9.25 9.22 10.92 13.51 11.70 D-Lyxose 9.50 13.08 10.64 9.87 12.65 16.05 13.96 13.02 15.35 18.33 16.33 Carbomix Ca-NP5 Maleic acid 8.42 9.50 8.53 8.56 / / / / / / / Mobile phase： H2O Malic acid 9.03 12.03 9.80 9.24 / / / / / / / Flow rate：0.6 mL/min Maltotetraose 7.42 / / / 7.78 / / / 9.40 12.11 10.31 Detector：RID Maltitol 8.04 10.51 8.41 8.29 10.77 14.74 11.92 11.24 16.69 22.68 18.44 Column Tem.： 85℃ D-(+)-Maltose 7.85 10.01 8.29 8.23 8.56 10.43 9.02 9.09 10.79 13.04 11.47 ： Maltotriose 7.54 8.94 7.70 7.90 8.08 9.40 8.33 8.57 10.23 12.49 10.87 Injection Volume 10 μL (3 injections) D-Mannitol 9.40 12.99 10.53 9.79 15.54 19.57 17.34 15.60 23.24 28.05 24.89 D-(+)-Mannose 9.11 12.55 10.13 9.48 11.00 14.09 12.05 11.45 15.01 18.43 16.09 Carbomix Pb-NP10 D-(+)-Melezitose 7.55 / / / 7.99 9.14 8.18 8.49 9.43 11.23 10.00 Mobile phase： H2O Methanol 17.99 20.60 19.21 18.28 15.32 17.74 16.08 15.64 15.82 17.19 16.31 Flow rate：0.6 mL/min Oxalic acid ： 7.61 7.72 7.44 7.72 / / / / / / / Detector RID dihydrate Column Tem.： 75℃ 1-Propanol 25.83 27.83 26.56 25.16 19.08 21.58 20.44 18.95 21.40 21.48 21.21 Injection Volume：10 μL (3 injections) 1,2-Propylene 15.43 / / / 16.23 / / / 19.68 20.88 20.10 glycol Carbomix Pb-NP5 Pyruvic acid 8.99 10.72 10.37 9.24 / / / / / / / ： D-(-)-Ribose 9.95 13.73 11.16 10.25 19.37 24.21 20.70 19.23 29.79 35.80 31.07 Mobile phase H2O D-Sorbitol 9.49 13.12 10.64 9.86 18.92 23.74 20.22 18.71 32.87 39.86 34.96 Flow rate：0.6 mL/min Succinic acid 10.46 14.45 11.54 10.47 / / / / / / / Detector：RID D-(+)-Sucrose / / / / 8.49 10.31 8.93 9.03 10.34 12.38 10.97 Column Tem.：75℃ Tartartic 8.41 10.74 8.94 8.64 / / / / / / / Injection Volume：10 μL (3 injections) Triethylene glycol 14.42 19.60 16.12 14.17 17.51 20.01 17.51 17.21 35.95 34.98 34.53 Xylitol 10.21 14.03 11.47 10.53 18.79 23.17 21.08 18.66 30.06 35.71 31.75 D-Xylose 9.19 12.61 10.24 9.60 10.72 13.49 11.63 11.19 12.93 15.24 13.70

www.sepax-tech.com Toll Free: 1-877-SEPAX-US CC1027 Mixed polysaccharides Analysis on Carbomix Pb-NP5:8% ( 7815 ) Column: Carbomix Pb-NP5:8% ( 5 µm, 8%, 7.8 x 150 mm ) Mobile phase: Water, Flow rate: 0.6 mL/min, Detector: RI ( 35 ℃ ) Column temperature: 80 ℃, Injection volume: 10 µL Samples: Cellobiose + Glucose + Mannose ( 5.0 mg/mL ), Pressure: 10 bar

nRIU

6.842

40000 6.056

8.223

30000

20000

10000

0 0 2 4 6 8 10 12 min

Cellobiose Glucose Mannose CC1027 Mixed polysaccharides Analysis on Carbomix Pb-NP5:8% ( 7815 ) Column: Carbomix Pb-NP5:8% ( 5 µm, 8%, 7.8 x 150 mm ) Mobile phase: Water, Flow rate: 0.6 mL/min, Detector: RI ( 35 ℃ ) Column temperature: 80 ℃, Injection volume: 10 µL Samples: Cellobiose + Glucose + Xylose + Arabinose ( 5.0 mg/mL ) Pressure: 10 bar

nRIU

8.110 6.841

7.268

40000 6.053

30000

20000

10000

0 0 2 4 6 8 10 12 min

Cellobiose Glucose Xylose Arabinose CC1027 Mixed polysaccharides Analysis on Carbomix Pb-NP5:8% ( 7815 ) Column: Carbomix Pb-NP5:8% ( 5 µm, 8%, 7.8 x 150 mm ) Mobile phase: Water, Flow rate: 0.6 mL/min, Detector: RI ( 35 ℃ ) Column temperature: 80 ℃, Injection volume: 10 µL Samples: Cellobiose + Glucose + Mannose + Xylose + Arabinose ( 5.0 mg/mL ) Pressure: 10 bar

nRIU

60000 Xylose

50000 Mannose Arabinose

Glucose Cellobiose

40000 30000 20000 10000 0 2 4 6 8 10 12 min

Cellobiose Glucose Xylose Arabinose/ Mannose CC1027 Mixed polysaccharides Analysis on Carbomix Pb-NP5:8% ( 7830 ) Column: Carbomix Pb-NP5:8% ( 5 µm, 8%, 7.8 x 300 mm ) Mobile phase: Water, Flow rate: 0.6 mL/min, Detector: RI ( 35 ℃ ), Column temperature: 80 ℃ Injection volume: 10 µL, Samples: Cellobiose + Glucose + Mannose ( 5.0 mg/mL ), Pressure: 27 bar

nRIU

25000 11.794 10.053

20000 15.066

15000

10000

5000

0 0 2.5 5 7.5 10 12.5 15 17.5 min

Cellobiose Glucose Mannose CC1027 Mixed polysaccharides Analysis on Carbomix Pb-NP5:8% ( 7830 ) Column: Carbomix Pb-NP5:8% ( 5 µm, 8%, 7.8 x 300 mm ) Mobile phase: Water, Flow rate: 0.6 mL/min, Detector: RI ( 35 ℃ ), Column temperature: 80 ℃ Injection volume: 10 µL, Samples: Cellobiose + Glucose + Xylose + Arabinose ( 5.0 mg/mL ) Pressure: 27 bar

nRIU

12.796 14.681

11.797

30000 Xylose

Glucose

Arabinose

10.056

Cellobiose

20000

10000

0 0 2 4 6 8 10 12 14 16 18 min

Cellobiose Glucose Xylose Arabinose CC1027 Mixed polysaccharides Analysis on Carbomix Pb-NP5:8% ( 7830 ) Column: Carbomix Pb-NP5:8% ( 5 µm, 8%, 7.8 x 300 mm ) Mobile phase: Water, Flow rate: 0.6 mL/min, Detector: RI ( 35 ℃ ), Column temperature: 80 ℃ Injection volume: 10 µL, Samples: Cellobiose + Glucose + Mannose + Xylose + Arabinose ( 5.0 mg/mL ) Pressure: 27 bar

nRIU

12.793 14.703

Xylose

Glucose

Mannose Arabinose

30000 Cellobiose

11.794

10.054 20000

10000

0 0 2 4 6 8 10 12 14 16 18 min

Cellobiose Glucose Xylose Arabinose /Mannose Tremella polysaccharide analysis on Zenix ( 7830 ) ZC1001 Column: Zenix-100 ( 3 µm, 100 Å, 7.8 x 300 mm), Zenix-80 ( 3 µm, 80 Å, 7.8 x 300 mm)

Mobile phase: 0.7% Na2SO4 , Flow rate: 0.5 mL/min Detector: RI ( 35℃ ), Column temperature: 35℃, Injection volume: 10 µL Sample: 5 mg/mL Tremella polysaccharide in mobile phase

nRIU 7000 10kD-100kD

6000

5000

4000

3000 Zenix-100

2000

1000 Zenix-80 0

0 5 10 15 20 25 min

www.sepax-tech.com 1 Better Surface Chemistry for Better Separation © Sepax Technologies, Inc. CC1001

Analysis of Carbohydrates on ® Carbomix Ca-NP5 (5 µm, 8%, 7.8x300 mm)

0.30

0.20 5 1

AU 6

4 0.10 2

0.00

-0.06

0 2 4 6 8 10 12 14 16 Min

Column: Carbomix® Ca-NP5 (5 µm, 8% crosslinkage, 7.8x300 mm) Mobile Phase: water Flow Rate: 0.6 mL/min Temperature: 85 oC Detection: UV 192 nm Injection Volume: 10 µL Samples (50 mM in 1. Melezitose 2. Maltose 3. Glucose 4. Mannose 5. Fructose H2O for each): 6. Adonitol (N=64,200/m)

Keywords: Carbohydrate, Carbomix, calcium form, sugars, carbohydrates

CC1003

Separation of Monosaccharide and its oligomers on ® Carbomix -Ca (5 µm, 8%, 4.6x300 mm)

G6-G5-G4-G3 -G2-G1

Column: Carbomix® Ca-NP5 (5 µm, 8% cross-linkage, 4.6x300 mm) Mobile Phase: water Flow Rate: 0.15 mL/min Temperature: 85 oC Detection: ELSD Injection Volume: 10 µL Samples: glucose (G1) and its oligomers (G2-G6) (Courtesy of Miyako Kawakatsu, M&S Instruments, Inc.)

Keywords: Carbohydrate, Carbomix, calcium form, sugar, glucose, monosaccharide, oligosaccharide

Analysis of Ethanol Production Broth on ® Carbomix -H (5 µm, 5%, 7.8 ID)

320.00 300.00 280.00 Maltotriose Ethanol ® 260.00 Carbomix -H (5 µm, Acetic acid 240.00 5%, 7.8 x 300 mm) 220.00 0.5 mL/min 200.00 180.00 Maltose 160.00 MV 140.00 120.00 100.00 Mathanol 80.00 Glucose 60.00 Stachyose 40.00 20.00 Glycerol 0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00 30.00 Minutes

Mobile Phase: 2.5 mM H2SO4 Temperature: 55 oC Flow Rate: 0.5 mL/min for 7.8 x 300 mm, 0.3 mL/min for 7.8 x 100 mm Detection: RI Injection volume: 20 µL Samples: stachyose, maltotriose, maltose, glucose, glycerol, acetic acid, methanol, ethanol

650.00 Maltotriose 600.00 ® Ethanol 550.00 Carbomix -H (5 µm, 5%, 7.8 x 100 Acetic acid 500.00 mm), 0.3 mL/min 450.00 400.00 Maltose 350.00

MV 300.00 250.00 200.00 Glucose Mathanol 150.00 Stachyose

100.00 Glycerol 50.00 0.00

1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00 15.00 16.00 17.00 18.00 19.00 20.00 Minutes

Keywords: Carbohydrate, Carbomix, hydrogen form, sugar, sugar alcohol, ethanol production broth, bio-fuel

CC1005

Analysis of Carbohydrates in Beer on ® Carbomix Ca (5 µm, 5%, 7.8x300 mm)

0.040

0.030

5 0.020

AU 0.010 1 2 3 4 0.000

-0.010

-0.020

2 4 6 8 10 12 14 16 18 20 22 Min

Column: Carbomix® Ca-NP5 (5 µm, 8% crosslinkage, 7.8x300 mm) Mobile Phase: water Temperature: 85 oC Flow Rate: 0.4 mL/min Detection: UV 192 nm Injection Volume: 2 µL Samples (6 mg/mL in H2O for each): 1. Maltotetraose, 2.Maltotriose, 3.Maltose, 4.Glucose, 5.Fructose.

Keywords: Carbohydrate, Carbomix, calcium form, beer, maltotetraose, maltotriose, maltose, glucose, fructose

CC1006

Analysis of Carbohydrates in Food on ® Carbomix -Ca (5 µm, 8%, 7.8x300 mm)

4 0.070

0.060

0.050 5

0.040 1

0.030 2 AU 3 0.020

0.010 0.000

-0.010

-0.020 2 4 6 8 10 12 14 16 18 22 22 24 26 Min

Column: Carbomix® Ca-NP5 (5 µm, 8% crosslinkage, 7.8x300 mm) Mobile Phase: water Temperature: 85 oC Flow Rate: 0.5 mL/min Detection: UV 192 nm Injection Volume: 2 µL Samples (6 mg/mL in H2O for each): 1. Sucrose, 2. Lactose, 3. Glucose, 4. Fructose, 5. Sorbitol.

0.30

0.20 5 1

AU 6

4 0.10 2

0.00

-0.06

0 2 4 6 8 10 12 14 16 Min

Column: Carbomix® Ca-NP5 (5 µm, 8% crosslinkage, 7.8x300 mm) Mobile phase: Water Flow rate: 0.6 mL/min Temperature: 85 °C Detection UV 192 nm Injection volume: 10 µL Samples (50 mM in 1. Melezitose 2. Maltose 3. Glucose 4. Mannose 5. Fructose H2O for each: 6. Adonitol (N=64,200/m)

Keywords: Carbohydrate, Carbomix, calcium form, food

Analysis of Carbohydrates on Carbomix® Ca-NP10 (10 µm, 8%, 7.8x300 mm)

Maltose

Xylose Lyxose

Ribose

2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00 Minutes

® Column: Carbomix Ca-NP10 (10 µm, 8%, 7.8x300 mm) Mobile Phase: water Temperature: 80 oC Flow Rate: 0.6 mL/min Detection: RI Samples: maltose, xylose, lyxose, ribose

Maltose Mannose

Fructose Glucose Melezitose Ribitol

2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 Minutes

Column: Carbomix® Ca-NP10 (10 µm, 8 %, 7.8x300 mm) Mobile phase: Water Temperature: 80 °C Flow rate: 0.5 mL/min Detection RI Injection Volume: 10 µL Samples: Melezitose, Maltose, Glucose, Mannose, Fructose, Ribitol

0 6 12 18

Minutes

Column: Carbomix® Ca-NP10 (10 µm, 8%, 7.8x300 mm) Mobile phase: Water Flow rate: 0.6 mL/min Temperature: 85 °C Detection: ELSD Injection volume: 0.5 µL Samples: Melezitose, Cellobiose, Glucose, Mannose, Fructose, Adonitol Lactose Galactitol Lyxose Galactose Ribose Glucose

0 6 12 18 24

Minutes

Column: Carbomix® Ca-NP10 (10 µm, 8%, 7.8x300 mm) Mobile Phase: water Flow Rate: 0.6 mL/min Temperature: 85 ºC Detection: ELSD Injection Volume: 2 µL Samples: Lactose, Glucose, Galactose, Lyxose, Galactitol, Ribose

Keywords: Carbohydrate, Carbomix, calcium form, xylose, ribose, melezitose, maltose, glucose, mannose, fructose, adonitol, cellobiose, lactose, galactose, lyxose, galactitol, ribitol

CC1008

Analysis of Sugar Mixture on Carbomix®-Pb (10 µm, 8%, 7.8x300 mm)

140.00

120.00

100.00 13.794

10.720 80.00 MV 14.688

60.00 12.718

16.158

34.450 40.00 38.253

20.00

0.00 0.00 5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 45.00 Minutes

Column: Carbomix Pb®-NP10 (10 m, 8% crosslinkage, 7.8x300 mm) Mobile Phase: water Temperature: 80 oC Flow Rate: 0.6 mL/min Detection: RI Sample Injection: 10 µL Samples: Cellobiose, Glucose, Xylose, Galactose, Arabinose, Xylitol, Sorbitol

Keywords: Carbohydrate, Carbomix, lead form, Xylose, Galactose, Arabinose

Analysis of Xylose, Galactose and Mannose on ® Carbomix -Pb (10 µm, 5%, 7.8x300 mm)

2 3

0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 Minutes

Column: Carbomix® Pb-NP10 (10 µm, 5% crosslinkage, 7.8x300 mm) Mobile Phase: water Temperature: 75 oC Flow Rate: 0.5 mL/min Detection: ELSD Sample Injection: 2 µL Samples: 1. Xylose, 2. Galactose, 3. Mannose

Keywords: Carbohydrate, Carbomix, lead form, xylose, galactose, mannose

Analysis of Lactose and Lactulose on Carbomix®-Pb (10 µm, 8%, 7.8x300 mm)

40.00

30.00 Lactose Lactulose

20.00

MV 10.00

0.00

2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 Minutes

Column: Carbomix®-Pb NP10 (10 µm, 8% crosslinkage, 7.8x300 mm) Mobile Phase: water Temperature: 75 oC Flow Rate: 0.6 mL/min Detection: RI Sample Injection: 5 µL Samples: Lactose & Lactulose

Keywords: Carbohydrate, Carbomix, lead form, lactose, lactulose

Analysis of Carbohydrates and Sugar Alcohols on Carbomix®-Pb (5 µm, 8%, 7.8x300 mm)

100.00 2 80.00

60.00 3 1 5

MV 4 6 40.00 7 8

20.00

0.00 0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00 30.00 32.00 34.00 36.00 38.00 40.00 42.00 44.00 Minutes

Column: Carbomix®-Pb (5 µm, 8% crosslinkage, 7.8x300 mm) Mobile Phase: water Temperature: 75 oC Flow Rate: 0.6 mL/min Detection: RI Sample Injection: 20 µL Samples: 1. Stachyose 2. Maltose 3.Glucose 4. Xylose 5. Galactose 6. Fructose 7. Mannitol 8. Sorbitol

700 Galactose Glucose 500 Fructose Mannitol mVolts 300 Sorbitol

100

0.0 10.0 20.0 30.0 40.0 50.0 Minutes

Column: Carbomix® Pb-NP5:8% (7.8*300 mm) Mobile Phase: H2O Flow Rate: 0.5 mL/min Temperature: 75 ºC Detection: ELSD Injection Volume: 5 µL Samples: glucose, galactose, fructose, mannitol, sorbitol

300 Xylose Cellobiose

200 Galactose mVolts Arabinose

100 Glucose Xylitol Sorbitol

0 0.0 10.0 20.0 30.0 40.0 50.0 Minutes Column: Carbomix® Pb-NP5:8% (7.8*300 mm) Mobile Phase: H2O Flow Rate: 0.5 mL/min Temperature: 75 ºC Detection: ELSD Injection Volume: 2 µL Samples: cellobiose, glucose, xylose, galactose, arabinose, xylitol, sorbitol

3 5 4

0 20 40 60

Minutes

Column: Carbomix® Pb-NP5:8% (7.8*300 mm) Mobile Phase: H2O Flow Rate: 0.5 mL/min Temperature: 75 ºC Detection: ELSD Injection Volume: 5 µL Samples: 1.Glucose; 2.Erythritol; 3.Mannitol; 4.Sorbitol; 5.Xylitol

Keywords: Carbohydrate, Carbomix, lead form, sugars, carbohydrates, sugar alcohols, stachyose, maltose, glucose, sylose, galactose, fructose, cellobiose, arabinose and erythritol, mannitol, sorbitol, xylitol

Analysis of Carbohydrates on Carbomix®-Na (5 µm, 8%, 7.8x300 mm)

30.00

25.00

20.00

MV 15.00

10.00

5.00

1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00 15.00 16.00

Minutes

Column: Carbomix® Na-NP5 (5 µm, 8% crosslinkage, 7.8x300 mm) Mobile Phase: water Temperature: 75 oC Flow Rate: 0.6 mL/min Detection: RI Sample Injection: 5 µL Samples: 1. Stachyose 2. Cellobiose 3.Glucose 4.Fructose 5. Arabinose 6. Ribose

Keywords: Carbohydrate, Carbomix, sodium form

Analysis of Carbohydrates on Carbomix®-K (5 µm, 8%, 7.8x300 mm)

160.00 140.00

120.00 100.00

80.00 MV

60.00

40.00

20.00

0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00 15.00 16.00 17.00 18.00 19.00 20.00 Minutes

Column: Carbomix® K-NP5 (5 µm, 8% crosslinkage, 7.8x300 mm) Mobile Phase: water Temperature: 85 oC Flow Rate: 0.4 mL/min Detection: RI Sample Injection: 5 µL Samples: 1. Stachyose 2. Cellobiose 3.Glucose 4.Fructose 5. Arabinose 6. Ribose

Keywords: Carbohydrate, Carbomix, potassium form

40 Melezitose

30 Cellobiose

mVolts 20

Mannose Adonitol

Fructose 10 Glucose

0 0 3 6 9 12 15 18 21 Minutes Column: Carbomix® Ca-NP10:8% (7.8*300 mm)

Mobile Phase: H2O Flow Rate: 0.6 mL/min

Temperature: 85 ºC

Detection: ELSD

Injection Volume: 0.5 µL

Samples: melezitose, cellobiose, glucose, mannose, fructose, adonitol

20 Lactose

10 mVolts Galactitol 5

0 Lyxose Galactose Ribose Glucose -5

0 6 12 18 24 Minutes Column: Carbomix® Ca-NP10 (10 µm, 8%, 7.8x300 mm)

Mobile Phase: H O 2 Flow Rate: 0.6 mL/min

Temperature: 85 ºC Detection: ELSD Injection Volume: 2 µL Samples: lactose, glucose, galactose, lyxose, galactitol, ribose

80 Erythritol

40 mVolts

20 Galactitol Arabinose Xylose Mannitol Glucose Melezitose Cellobiose Adonitol 0

0 6 12 18 24 Minutes ® Column: Carbomix Ca-NP10 (10 µm, 8%, 7.8x300 mm)

Mobile Phase: H O 2 Flow Rate: 0.6 mL/min

Temperature: 85 ºC

Detection: ELSD

Injection Volume: 5 µL

Samples: melezitose, cellobiose, glucose, xylose, arabinose, erythritol, mannitol, galactitol

5 Erythritol Maltose Maltitol mVolts 0 Maltotriose Fructose Sorbitol Glucose

-5

0 6 12 18 24

Minutes Column: Carbomix® Ca-NP10 (10 µm, 8%, 7.8x300 mm) Mobile Phase: H2O Flow Rate: 0.6 mL/min Temperature: 85 ºC Detection: ELSD Injection Volume: 2 µL Samples: maltotriose, maltose, glucose, maltitol, fructose, erythritol, sorbitol

100 Lactose

60 Mannose Galactitol

mVolts 40

20 Lyxose Ribose

0 10 20 30 Minutes Column: Carbomix® Ca-NP10 (10 µm, 8%, 7.8x300 mm) Mobile Phase: H2O Flow Rate: 0.6 mL/min Temperature: 85 ºC Detection: ELSD Injection Volume: 5 µL Samples: lactose, mannose, lyxose, galactitol, ribose

200 Sorbitol

150

100 mVolts

50 Mannitol

0 0 6 12 18 24 30 Minutes Column: Carbomix® Ca-NP10 (10 µm, 8%, 7.8x300 mm) Mobile Phase: H2O Flow Rate: 0.6 mL/min Temperature: 85 ºC Detection: ELSD Injection Volume: 1 µL Samples: mannitol, sorbitol

Keywords: Carbohydrate, Carbomix, calcium form, ELSD, sugar mixture, sugar alcohols

CC1014a

Analysis of Sugar on Carbomix® Ca-NP10 (10 µm, 8%, 7.8x300 mm) - Comparison of the two detectors: ELSD and RID

8 80

60 6

5 mM, 2 µL@ELSD mVolts

mVolts 40 4

20 5 mM, 10 µL@ ELSD

2 5 mM, 10 µL@ RID

0 0 4 8 12 16 Minutes

Column: Carbomix® Ca-NP10 (10 µm, 8% crosslinking, 7.8x300 mm) 1. ELSD 2. RID Mobile Phase: H2O H2O Flow Rate: 0.6 mL/min 0.6 mL/min Temperature: 85 OC 85 OC Detection: ELSD (drift tube temp.: 65 OC; RI (temp.: 30 OC gas flow rate: 2.0 L/min Injection Volume: 10 µL 10 µL Samples: Glucose and Fructose, 5 mM Glucose and Fructose, 5 mM

Keywords: Carbohydrate, Carbomix, calcium form, glucose, fructose, ELSD, RI

CC1015

Analysis of sugar and sugar alcohol on Carbomix® NP10 (10 µm, 8%, 7.8x300 mm) ® ® - Comparison of Carbomix H and Carbomix Ca

1 2

Column: Carbomix H-NP10:8%(7.8*300)

Column: Carbomix Ca-NP10:8%(7.8*300)

0 6 12 18 24 Minutes

Columns: Carbomix® H-NP10:8% and Carbomix® Ca-NP10 (10 µm, 8% crosslinking, 7.8x300 mm) ® ® Mobile Phases: TFA solution, pH 2.5 for Carbomix H; H2O for Carbomix Ca Flow Rate: 0.6 mL/min Detection: ELSD Samples: 1. xylose; 2. xylitol

Column: Carbomix H-NP10:8%(7.8*300)

Column: Carbomix Ca-NP10:8%(7.8*300)

0 4 8 12 16 Minutes

Columns: Carbomix® H-NP10 and Carbomix® Ca-NP10 (10 µm, 8% crosslinking, 7.8x300 mm) ® ® Mobile phase: TFA solution, pH 2.5 for Carbomix H; H2O for Carbomix Ca Flow rate: 0.06 mL/min Detection: ELSD Samples: 1. Maltose 2. Maltitol

Keywords: Carbohydrate, Carbomix, ELSD, sugar and sugar alcohol, xylose and xylitol, maltose and maltitol

CC1016

Analysis of glucose and fructose on Carbomix® H/Ca/Pb (7.8x300 mm)

2 1

Column: Carbomix H-NP10:8%(7.8*300)

Column: Carbomix Ca-NP10:8%(7.8*300)

1 2

Column: Carbomix Pb-NP5:8%(7.8*300)

0.0 10.0 20.0 30.0 40.0 50.0 Minutes

Columns: Carbomix® H-NP10; Carbomix® Ca-NP10; Carbomix® Pb-NP5: (8% crosslinking, 7.8x300 mm) ® ® Mobile Phases: TFA solution, pH 2.5 for Carbomix H; H2O for Carbomix Ca and Carbomix® Pb Flow Rate: 0.6 mL/min Detection: ELSD Samples: 1. glucose; 2. fructose

Keywords: Carbohydrate, Carbomix, ELSD, glucose and fructose, ionic form

CC1017

Analysis of orange juice on Carbomix® Ca-NP10 (10 µm, 8%, 7.8x300 mm)

Fructose

Glucose

0 4 8 12 16

Minutes

Column: Carbomix® Ca-NP10 (10 µm, 8%, 7.8x300 mm) Mobile Phase: H2O Flow Rate: 0.6 mL/min Temperature: 85 ºC Detection: ELSD Injection Volume: 10 µL Samples: glucose and fructose, 5 mM

Keywords: Carbohydrate, Carbomix, calcium form, ELSD, orange juice

CC1018

Analysis of Carbohydrates in Orange Juice on Carbomix®-Ca (5 µm, 8%) - Fast analysis with shorter column

1 1. Sucrose 2. Glucose 1 3. Fructose

2 3

(a) 2

(b)

0.0 2.0 4.0 6.0 8.0 10.0 12.0 14.0 16.0 18.0 20.0 Min

Columns: Carbomix® Ca-NP5 (5 µm, 8% crosslinkage) (a) 7.8 x 100 mm; (b) 7.8 x 300 mm Mobile Phase: water Temperature: 85 oC Flow Rate: 0.6 mL/min Detection: RI Injection Volume: 10 µL Samples: Sucrose, glucose and fructose (15 mM/each)

Keywords: Carbohydrate, Carbomix, calcium form, sugar, orange juice, sucrose, glucose, fructose

CC1019 Analysis of sugar mixture on Carbomix® H-NP10 (10 µm, 8%, 7.8x300 mm) mm) 60

Fructose

40 Glucose

30 mVolts

20 Sucrose

0 0 2 4 6 8 10 12 14 Minutes Column: Carbomix® H-NP10 (10 µm, 8%, 7.8x300 mm) Mobile Phase: TFA solution, pH 2.5 Flow Rate: 0.6 mL/min Temperature: 55 ºC Detection: ELSD Injection Volume: 10 µL Samples: sucrose, glucose, fructose

600 Lactose

400 mVolts Mannose 200 Ribose

0 0 4 8 12 16 Minutes Column: Carbomix® H-NP10 (10 µm, 8%, 7.8x300 mm)

Mobile Phase: TFA solution, pH 2.5

Flow Rate: 0.6 mL/min

Temperature: 55 ºC

Detection: ELSD

Injection Volume: 5 µL Samples: lactose, mannose, ribose

500 Maltose

400

300 Succinic acid/Erythritol Succinic mVolts 200 Adonitol Glucose 100 Tartaric Lyxose Maltotriose Galactose

0 0 4 8 12 16 Minutes Column: Carbomix® H-NP10 (10 µm, 8%, 7.8x300 mm)

Mobile Phase: TFA solution, pH 2.5

Flow Rate: 0.6 mL/min

Temperature: 55 ºC

Detection: ELSD

Injection Volume: 10 µL

Samples: maltotriose, maltose, tartaric, glucose, galactose, lyxose, adonitol, succinic acid, erythritol

400 Sorbitol mVolts 200 Xylose Xylitol Arabinose Glucose

0 4 8 12 16

Minutes ® Column: Carbomix H-NP10 (10 µm, 8%, 7.8*300 mm)

Mobile Phase: TFA solution, pH 2.5 Flow Rate: 0.6 mL/min Temperature: 55 ºC Detection: ELSD Injection Volume: 10 µL Samples: glucose, xylose, sorbitol, arabinose, xylitol

Keywords: Carbohydrate, Carbomix, hydrogen form, ELSD, sugar mixture

CC1020 Analysis of sugar mixture on Carbomix®-Pb (10 µm, 5%, 7.8x300 mm)

Column: ® Carbomix Pb -NP10 (10 m, 5% crosslinkage, 7.8x300 mm) Mobile Phase: water

Temperature: 75 oC Flow Rate: 0.5 mL/min Detection: ELSD: drift tube temp.: 80 OC; gas flow rate: 2.0 L/min Injection volume: 5 µL

Column: ® Carbomix Pb -NP10 (10 m, 5% crosslinkage, 7.8x300 mm) Mobile Phase: water

Temperature: 75 oC Flow Rate: 0.3 mL/min Detection: ELSD: drift tube temp.: 80 OC; gas flow rate: 2.0 L/min Injection volume: 5 µL

18.0

15.0 Sucrose Sucrose

12.0 Glucose MV

9.0 Xylose

6.0 Galactose 43.597 Arabinose Fructose 3.0

0.0 10.0 20.0 30.0 40.0 50.0 Minutes

® Column: Carbomix Pb -NP10 (10 m, 5% crosslinkage, 7.8x300 mm) Mobile Phase: water Temperature: 75 oC Flow Rate: 0.5 mL/min Detection: RI Injection volume: 20 µL

Keywords: Carbohydrate, Carbomix, lead form, Sucrose, Glucose, Xylose, Galactose, Arabinose, mannose, fructose

® Analysis of Maltotriose and Maltotetraose on Carbomix H-NP10 (10 µm, 5%, 7.8x300 mm)

28.00 26.00 24.00 22.00 20.00 18.00 Maltotetraose Maltotriose 16.00 14.00 12.00

10.00MV 8.00 6.00 4.00 2.00 0.00 -2.00 -4.00 -6.00 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00 10.50 11.00 11.50 12.00 12.50 13.00 Minutes

Column: Carbomix® H-NP10:5% (7.8*300 mm) Mobile Phase: 2.5 mM H2SO4 Flow Rate: 0.5 mL/min Temperature: 55 ºC Detection: RI Injection Volume: 20 µL Samples: Maltotiose and Maltotetraose (0.1, 0.2, 0.4, 0.6 and 0.8 mg/ml in water)

Calibration standard curve on Page 2

Maltotetraose/(mg/mL) 0.0 0.1 0.2 0.4 0.6 0.8 1.0 Area 0 30677 70406 144197 217613 292733 356482

Maltotriose/(mg/mL) 0.0 0.1 0.2 0.4 0.6 0.8 1.0 Area 0 30614 74714 148368 223655 308321 383919

400000 y = 360210x 500000 R2= 0.9994 300000 400000 y = 381050x R2= 0.999 300000 200000 Area Area 200000 100000 100000

0 0 0 0.2 0.4 0.6 0.8 1 1.2 0 0.2 0.4 0.6 0.8 1 1.2 Maltotetraose/(mg/mL) Maltotriose/(mg/mL)

Keywords: Carbohydrate, Carbomix, hydrogen form, RI, sugar mixture, maltotetraose, maltotriose

CC1022

® Loading Capacity Test of Carbomix -Ca NP10:8% (10 µm, 8%, 7.8x300 mm) With Glucose and Fructose

4000

3500 Glucose Fructose

3000

2500

2000 3600 g MV 3150 g 1500 2700 g 2250 g 1000 1800 g 1620 g 500 1260 g 900 g 0 0.0 5.0 10.0 15.0 20.0 Minutes

Column: Carbomix® Ca-NP10:8% (7.8x300 mm) Mobile Phase: H2O Flow Rate: 0.6 mL/min Temperature: 85 ºC Detection: RI Injection amount: 0.9, 1.26, 1.62, 1.8, 2.25, 2.7, 3.15 and 3.6 mg each Samples: Glucose and Fructose

Keyword: Carbomix, Carbomix Ca-NP10:8%, capacity, carbohydrate, glucose, fructose

Analysis of sugar and sugar alcohol mixture on Carbomix® H/Ca/Pb (7.8x300 mm)

3,4

3 4,5 Column: Carbomix H-NP10:8% Column: Carbomix Ca-NP10:8% 2 (7.8 x 300 mm) (7.8 x 300 mm) Mobile phase: TFA, pH 2.5 Mobile phase: water Flow rate: 0.6 mL/min Flow rate: 0.6 mL/min Temp erature: 55 ºC Temp erature: 75 ºC 1 5 Detector: ELSD Detector: ELSD 2 1

4 8 12 16 0 8 16 24 Minutes Minutes 2 Column: Carbomix Pb-NP10:8% Column: Carbomix Pb-NP5:8% 3 (7.8 x 300 mm) (7.8 x 300 mm) 3 Mobile phase: water 2 Mobile phase: water 1 Flow rate: 0.6 mL/min Flow rate: 0.5 mL/min 1 Temp erature: 85 ºC Temp erature: 75 ºC 5 4 Detector: ELSD 5 4 Detector: ELSD

0 20 40 60 0 20 40 60 Minutes Minutes

Conditions indicated on Figures Samples: 1. glucose; 2. Erythritol; 3. Mannitol; 4. Sorbitol; 5. Xylitol

Keywords : Carbohydrate, Carbomix, ELSD, glucose, erythritol, mannitol, sorbitol, xylitol, ionic form

SL1002

Separation of Low MW Polysaccharides on SRT®

SRT SEC- -100 (100 Å)

SRT SEC-150 (150 Å)

0.0 2.0 4.0 6.0 8.0 10.0 12.0 Min

Column: SRT® (5 µm, 4.6 x 300 mm) Mobile Phase: 0.15 M phosphate buffer, pH 7 Flow Rate: 0.35 mL/min Detection: UV 214 nm Sample: Achyranthes bidentata polysaccharides (ABPS) from plant root extract(MW < 10,000) Injection Volume: 10 µL

Keywords: size exclusion, SRT, polysaccharides

Separation of Carbohydrates on Proteomix® SAX-NP5 (5m, 4.6x150 mm)

Starch hydrolysate (5 µL)

Glucose oligomer (G1-G6) (5 µL)

Column: Proteomix® SAX-NP5 (5m, 4.6x150mm) Mobile Phases: (A) 0.05% (25% NH4OH) in CH3CN; (B) 0.05% (25% NH4OH) in H2O Gradient: 15-80%B (20 min) Flow Rate: 0.7 mL/min Temperature: 35 ºC Detection: ELSD Samples: Corn starch hydrolysate; glucose oligomer (G1 to G6) (Courtesy of Miyako Kawakatsu, M&S Instruments Inc.)

Keywords: Ion-exchange, Proteomix, strong anion exchange, carbohydrate, corn starch hydrolysate, glucose oligomer, volatile buffer

Analysis of Xylo-oligosaccharide on Proteomix® SAX-NP5 (5 µm, 4.6x150 mm)

600

400

mVolts

200

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Minutes

Column: Proteomix® SAX-NP5 (5 µm, 4.6x150 mm) Mobile Phases: A: 0.05% NH4OH in ACN; B: 0.05% NH4OH in H2O Gradient: 15-60%B (20 min) Flow Rate: 0.7 mL/min Back Pressure: 70 Bar Detection: ELSD Temperature: 35 ºC Injection Volume: 10 μL Sample: Xylose oligosaccharides

Keywords: Ion-exchange, Proteomix, strong anion exchange, xylose oligosaccharides

Separation of Glucose/Xylose/Galactose/L-Arabinose/Mannose/Fructose on two Carbomix® Pb-NP10 (10 µm, 5%, 7830) in tandem

100.00

90.00

80.00 Glucose

70.00 Galactose Xylose Arabinose 60.00 - L Mannose Fructose 50.00 MV 40.00

30.00

20.00

10.00

0.00

0.0 17.0 34.0 51.0 68.0 85.0 Minutes

Column: 2 x Carbomix® Pb-NP10 (10 µm, 5% crosslinkage, 7.8x300 mm)

Mobile Phase: water o Temperature: 75 C Flow Rate: 0.3 mL/min Detection: RI Sample Injection: 10 µL Samples: Glucose/Xylose/Galactose/L-Arabinose/Mannose/Fructose (0.5 M each)

Keywords: Carbohydrate, Carbomix, lead form, glucose, mannose, arabinose, xylose, galactose, fructose

® Column Temperature Effect on Sugar Separations on Carbomix Ca-NP5:8% (5 µm, 8%, 7.8x300 mm)

nRIU 30000 60oC 25000 1. Sucrose 1 20000 2. Glucose 15000 4 3. Xylose 2 3 10000 4. Fructose 5000 0 nRIU 30000 1 o 2 25000 80 C 3 4 20000 15000 10000 5000 0 4 6 8 10 12 14 16 18 min

Column: Carbomix® Ca-NP5:8% (5 µm, 8% crosslinking, 7.8x300 mm)

Mobile Phase: water o o Temperature: 60 C, 80 C Flow Rate: 0.5 mL/min Detection: RI Injection volume: 2.5 µL Samples: Sucrose/Glucose/Xylose /Fructose (20 mg/mL each)

Keywords: Carbohydrate, Carbomix, Calcium form, NP5, 8% crosslinking, glucose, sucrose, xylose, fructose, column temperature, 60 oC, 80 oC